Minimal residual disease testing by flow cytometry for childhood Precursor B cell Acute Lymphoblastic Leukaemia – Royal Children’s Hospital experience.

The presence of MRD at day 29 of treatment for precursor B cell Acute Lymphocytic Leukaemia (Pre B ALL) is the single most important prognostic factor in children treated on the Children’s Oncology Group (COG) protocols.1 MRD assessment is now considered standard of care in the treatment of children with ALL.

The Cell Therapy and Flow Cytometry Laboratory at the Royal Children’s Hospital, Melbourne have performed Minimal Residual Disease (MRD) testing by flow cytometry since 2011, using protocols identical to the COG method. In excess of 230 patients have been monitored with >700 tests (including external referrals). The assay examines differences from normal expression of CD9, CD10, CD13/33, CD19, CD20, CD34, CD38, CD45 and CD58. Clusters of events that fall outside the areas where normal cells occupy are considered as abnormal. Using these markers 93% of the patients had an informative phenotype. 1 x 106 cells are labelled in order to collect enough events to give a sensitivity of at least 0.01%. Our results compare well with molecular MRD testing by Ig/TCR gene rearrangements.2 7 patients that have been monitored since diagnosis have undergone allogeneic stem cell transplant and the MRD panel has also been used to assess post –transplant MRD and to tailor immunosuppression where appropriate.

MRD detection by flow cytometry has the advantages of being a rapid test, which is applicable to almost all Pre B ALL patients. It is the standard of care for the treatment of all our paediatric patients with Pre B ALL.

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